Induction of c-fos gene by mercury chloride in LLC-PK1 cells.
The c-fos, a member of the immediate early genes, has been reported to be expressed in the renal proximal tubule in response to ischemic and toxic injury. In the present study, effects of mercury chloride (HgCl2) on the expression of c-fos were examined in LLC-PK1 cells. The reverse transcription polymerase chain reaction (RT-PCR) analysis for the semi-quantification of mRNA showed that the treatment of 20 microM HgCl2, markedly increased c-fos mRNA levels. The level of c-fos mRNA began to increase after a 30-min exposure, peaked at 1 h and then returned to the control level at 8 h. The HgCl2-induced c-fos expression was abolished completely by actinomycin-D, indicating it was due to transcriptional activation of the gene. Western blotting immunodetection revealed accumulation of c-Fos protein after 1 h exposure to 20 microM HgCl2. The cytotoxicity of HgCl2 as assayed by mitochondrial dehydrogenase activity (MTT conversion) was observed after 18 h exposure but not at 0.5-8 h. Also, the decrease in cell viability was accompanied with DNA fragmentation, which is characteristic of apoptosis. The present results showed that HgCl2 could induce the early expression of c-fos gene in a renal epithelial cell line.
Matsuoka
M
M
Department of Environmental Toxicology, University of Occupational and Environmental Health, Kitakyyushu, Japan. masatomm@med.uoehu.ac.jp
Wispriyono
B
B
Igisu
H
H
D000818:Animals; D015153:Blotting, Western; D002470:Cell Survival / Q000187:drug effects; D004249:DNA Damage / Q000187:drug effects; D017931:DNA Primers / Q000737:chemistry; D003609:Dactinomycin / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D016762:Genes, fos / Q000235:genetics*; D018374:LLC-PK1 Cells / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D008627:Mercuric Chloride / Q000633:toxicity*; D009707:Nucleosomes / Q000187:drug effects; D016133:Polymerase Chain Reaction; D016760:Proto-Oncogene Proteins c-fos / Q000096:biosynthesis* / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis; D013552:Swine
D017931:DNA Primers; D009707:Nucleosomes; D016760:Proto-Oncogene Proteins c-fos; D012333:RNA, Messenger; D003609:Dactinomycin; D008627:Mercuric Chloride